• Journal of Life Science
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• v.28 no.10
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• pp.1201-1211
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• 2018

The study compared the anti-aging, anti-adipogenesis, and anti-tumor effects of epigallocatechin-3- gallate (EGCG) in various cancer cell lines (SNU-601, MKN74, AGS, MCF-7, U87-MG, and A-549) and normal cell lines (MRC-5 fibroblasts, dental tissue-derived mesenchymal stem cells [DSC], and 3T3-L1 pro-adipocytes). Half inhibitory concentration ($IC_{50}$) values were significantly (p<0.05) higher in normal cell lines (~50 uM), when compared to that in cancer cell lines (~10 uM). For anti-aging effects, MRC-5 and DSC were exposed to 10 uM EGCG for up to five passages that did not display any growth arrest. Population doubling time and senescence-related ${\beta}-galactosidase$ ($SA-{\beta}-gal$) activity in treated cells were similar to untreated cells. For anti-adipogenic effects, mouse 3T3-L1 pre-adipocytes were induced to adipocytes in an adipogenic differentiation medium containing 10 uM EGCG, but adipogenesis in 3T3-L1 cells was not inhibited by EGCG treatment. For anti-tumor effects, the cancer cell lines were treated with 10 uM EGCG. PDT was significantly (p<0.05) increased in EGCG-treated SNU-601, AGS, MCF-7, and U87-MG cancer cell lines, except in MKN74 and A-549. The level of telomerase activity and cell migration capacity were significantly (p<0.05) reduced, while $SA-{\beta}-gal$ activity was highly up-regulated in EGCG treated-cancer cell lines, when compared to that in untreated cancer cell lines. Our results have demonstrated that EGCG treatment induces anti-tumor effects more efficiently as noted by decreased cell proliferation, cell migration, telomerase activity, and increased $SA-{\beta}-gal$ activity than inducing anti-aging and anti-adipogenesis. Therefore, EGCG at a specific concentration can be considered for a potential anti-tumor drug.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.98-98
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• 1995

독사 또는 곤충의 독 30여종을 대상으로 SNU-1 위암세포에 대하여 MTT test를 실시한 결과 세포 독성 활성이 제일 높은 킹코브라(Ophiophagus hannah)의 venom을 가지고 세포 독성 물질을 정제하였다. Gel Filtration Chromatography와 Anion Exchange Chromatography로 정제한 4번째 peak만이 MTT/SRB test결과 IC$_{50}$ value 0.947$\mu\textrm{g}$/ml이었다. 음이온 교환 크로마토그라피로 정제한 단백질을 PRO-RPC로 더 분리하여 순수한 단일성분을 얻었으며 맹장암, 대장암, melanoma, fibrosarcoma 세포에 대해 독성을 확인하였고, 광학 및 전자 현미경에 의해 암세포의 분화와 성장이 억제됨을 재확인하였다. 또한 thymidine uptake asaay에서 암세포의 증식이 억제되었고, 또한 EDTA, $Zn^{++}$, $Ba^{++}$ 첨가로 세포 독성 활성이 증가되었다. (중략)

• Microbiology and Biotechnology Letters
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• v.34 no.2
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• pp.143-149
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• 2006

A extract from Spirulina platensis of seawater and freshwater was obtained by using the water and ethanol. Extraction yields of seawater S. platensis were observed about 3% higher than freshwater S. platensis. Cytotoxicity (HEK293) and inhibition ratio of cancer cell line (A549, AGS, MCF7, Hep3B) in adding of the extracts from the S. platensis of seawater and freshwater were measured by SRB assay. Cytotoxicity of all of the extracts in adding 1.0 mg/ml was below 26%. Ctotoxicity of the extracts from the seawater S. platensis were about 6% less than freshwater S. platensis. Inhibition ratio of cancer cell growth was inhibited by adding 1.0 mg/ml of the extracts that was obtained about 80%. Inhibition effect of cancer cell growth in adding seawater S. platensis was observed higher than freshwater S. platensis. Differentiation ratio of HL-60 cells in adding the extracts of seawater S. platensis was observed highly that was 160.9%.

• Journal of Life Science
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• v.21 no.4
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• pp.529-533
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• 2011

Metallothioneins (MT) are a group of low-molecular weight, cysteine-rich, intracellular proteins that are encoded by a family of genes containing at least 10 functional isoforms in human. The expression and induction of these proteins is associated with protection against DNA damage, oxidative stress, and apoptosis. Many studies have shown increased expression of MT in various human tumors, whereas MT is down-regulated in certain tumors such as hepatocellular carcinoma and liver adenocarcinoma. Hence, the expression of MT is not universal to all human tumors but may depend on the differentiation status and proliferative index of tumors, along with other tissue factors and gene mutations. Using Northern blot analysis, we found that laminin induced expression of MT-1 in HSG and PC12 cells, which can be differentiated by laminin, but had no effect on MB-231, MDA-435, and PC-3 cells, which cannot be differentiated by laminin. In addition, we analyzed the expression level of the MT-1 gene in five prostate cancer cell lines possessing different metastatic potential. The expression of MT-1 in normal and less malignant cells (RWPE-1 and WPE1-NA22) was high and up-regulated by laminin, whereas the expression of MT-1 in WPE1-NB14, WPE1-NB11, and WPE1-NB26 cells (malignant) was extremely low and not elevated by laminin. These results suggest that the MT-1 gene is involved in laminin-mediated differentiation and affects the metastatic potential of tumor cells.

• Journal of Life Science
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• v.33 no.9
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• pp.693-702
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• 2023

The present study examined the rate of cell growth and differentiation potential into adipocytes in A549 lung adenocarcinoma cells exposed to each adipogenic medium containing glucose metabolism hormones, such as thyroxine (T4) thyroid hormone and glucocorticoid (GC) adrenal steroid hormone, as well as pioglitazone (PGZ), a PPARγ agonist. Following each adipogenic treatment for 2 weeks, the rate of cell growth was significantly (p<0.05) inhibited, and the level of telomerase activity was significantly (p<0.05) decreased in the PGZ-based adipogenic medium containing both T4 and GC hormone compared with those containing each T4 or GC hormone. Moreover, the adiposome-like vesicles were highly reacted with Oil-Red O staining solution, and the levels of transcripts expressed in the differentiating adipocytes for adipogenesis, including adinopectin, leptin, and resistin, were significantly (p<0.05) increased in the PGZ-based adipogenic medium containing both T4 and GC hormone compared with those of the adipogenic medium containing each T4 or GC hormone, implying that adipocytic differentiation has fully occurred in the A549 cancer cells. Based on present observations, the PGZ-based adipogenic medium containing both T4 and GC efficiently induces inhibition of cell growth and cellular differentiation into adipocytes in A549 cancer cells rather than in the adipogenic medium containing only T4 or GC hormone. Adipogenic treatment could provide potential probability in cancer chemotherapy.

• The Korean Journal of Zoology
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• v.37 no.2
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• pp.182-202
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• 1994

마우스(ICR mouse)의 줄생전 ·후 발생에 따른 설점막에서 설유두와 미뢰의 발생, 미뢰를 구성하는 미각세포들의 각 형별 미세구조적인 특징 및 미각세포와 신경종말과의 관계를 광학 및 전자현미경으로 관찰하였다. 유곽유두는 임신 17일 심상유두와 엽상유두는 출생 후 0일, 사상유두는 출생후 1일에 출현하였으며, 유곽유두와 심상유두에서의 미뢰는 출생 후 0일, 엽상유두에서의 미뢰는 출생후 1일에 관찰되었다. 유곽유두에서는 유두벽 상피 뿐만 아니라 유두 표면상피에서도 성체에서와는 다르게 미뢰가 출현하였다. 출생후 7일과 15일에서 유곽유두의 미뢰에 대한 미세구조적 관찰에서 미각세포는 중간형 세포. 암세포, 명세포, 기저세포 및 주변세포로 구분되었다 중간형 세포는 암세포와 명세포의 중간정도의 전자밀도를 보이는 세포로서 세포질에서는 활면 및 조면소포체가 다수 관찰되었고. 세포질 및 핵질내에서 다소포성소체와 유사한 구조물들이 관찰되었다. 암세포는 전자밀도가 높아 어둡게 보였고. 핵막은 깊은 함입을 보였다 이 세포의 핵주변 세포질에는 미토콘드리아, 섬유상 물질, 조면소포체 및 폴리솜들이 다수 관찰되었으며, 첨부 세포질에서는 전자밀도가 높아 어둡게 보이는 과립들이 다수 관찰되었다. 명세포는 전자밀도가 낮아 밝게 보였고, 크고 등근 핵을 갖고 있었으며. 세포질내에서 활면소포체, 폴리솜 그리고 크고 작은 공포들이 다수, 조면소포체들은 소수 관찰되었다. 기저세포는 핵질이 풍부한 난원형의 핵을 갖고 있었으며, 전자밀도가 낮아 밝게 보였고, 유리리보솜들은 많았으나, 활면 및 조면소포체들은 소수 관찰되었다 주변세포는 비교적 긴 방추형의 세포로 얕은 핵막 함입들을 보였고, 대부분의 세포소기 관들은 미약하였으나 유리리보솜들은 다소 많이 관찰되었다. 출생후 7일에는 명세포, 15일에는 중간형 세포들이 다른 세포에 비하여 다수 출현하였다 출생후 미각세포들의 형태적 특징 및 세포소기 관들의 분화들로 미루어 암세포 주변세포 및 기저세포는 미성숙세포, 중간형 세포는 미각기능이 활발한 성숙한 세포 그리고 명세포는 퇴행성 세포로 생각된다.

• Proceedings of the Korean Vacuum Society Conference
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• 2012.02a
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• pp.558-558
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• 2012

최근의 원자간력현미경(AFM)은 soft한 생체물질을 비파괴적 방법 및 나노크기의 분해능으로 여러 구조적, 물리적 특성 측정이 가능하여 bio분야에 다양이 활용되고 있다. 본 연구에서는 AFM을 이용하여 줄기세포인 BM MSC(bone marrow mesenchymal stem cell)가 신경세포로 분화 여부를 측정하는 방법을 보고하고자 한다. 신경세포의 신호전달은 시냅스에서 신경전달물질을 매개로 하여 이루어지는데, 신경전달물질 중에 D-Glutamic acid는 시냅스후세포에서 흥분성 전위 크기를 증가시킨 상태를 장기간 유지시켜주는 물질로, 특정물질인 Glutamate와 항원-항체 결합을 한다. 본 연구에서는 이 두 물질간의 항원-항체 반응을 활용하여 줄기세포의 신경세포로 분화 여부를 AFM으로 측정하였다. 먼저, 수용성 시료인 두 물질을 증류수에 용해시켜 Mica 기판에 그 용액을 떨어뜨려 자연건조로 시료를 준비한 후, AFM으로 형태 및 크기를 측정하였다. D-Glutamic acid와 Glutamate는 구형 입자 형태를 보였으며, Glutamate의 너비는 ~100 nm이고, D-Glutamic acid는 ~50 nm였다. 두 물질이 든 용액을 섞었을 때, 항원-항체 반응에 의해 다른 크기의 두 구형입자가 붙어 있는 형태가 관찰되었다. 이 반응을 활용하여, 신경세포에서 분비되는 신경전달물질인 D-Glutamic acid를 선별하였다. DMEM 배지에 신경암세포주인 SH-SY5Y 를 접종한 후 $37.6^{\circ}C$의 incubator에서 24시간 배양하고, 화학적 자극(60~70 mM의 KCl 용액을 주입함)을 주어 신경전달물질 분비를 유도하였다. 그 배지에 항체 Glutamate 를 주입하여 자연건조 시킨 후 항원-항체 결합특성을 AFM으로 측정하여, 항원-항체 결합된 이미지와 동일함을 확인하였다. 결과적으로 AFM을 이용한 신경전달물질의 항원-항체 결합여부 측정을 통해, BM MSC 줄기세포의 신경세포로 분화를 판단할 수 있으며, 이 방법은 줄기세포의 특정 세포로의 분화 여부 판단에 활용될 것으로 기대된다.

• Applied Microscopy
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• v.29 no.1
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• pp.105-124
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• 1999

Histological and ultrastructural differentiations of the neuroepithelial cells in the mouse embryo during neurulation were observed. The neural plates and grooves consisted of pseudostratified columnar epithelium in the embryonic day (ED) 8 embryo were developed. In the ED 9 embryo, the neural tube was developed in all body length of embryo except both the cephalic and caudal ends. Secondary neurulation was shown at the tail bud of the ED 10 embryo. In the ED 8 embryo, the primitive streak was shown in the posterior end of the embryonic disc. The neuroepithelium, notochord and mesenchyme were well differentiated in the cephalic and cervical portions. In the ED 9 and 10 embryos, the roof plates of neural tubes were constituted of the closing of the surface ectodermal cells in the hindbrain and the neuroepithelial cells in the spinal cord. The floor plate of neural tube were consisted of the low pseudostratified columnar epithelium. The spinal motor nerve fibers were initially differentiated in the ED 10 embryo. According to the electron density of the cell and the differentiation of tell organelles, the neuroepithelial cells in the ED 9 and 10 embryos were classified into three types: dark, intermediate and light types. All types in the ED 9 embryo were observed but the dark cell in the ED 10 embryo was not done. The free ribosomes and polysomes in all neuroepithelial cells were developed. The RER and lipid droplets in the dark cell and the Golgi complex in the intermediate and light cells were observed. Many microfilaments in the cytoplasmic processes of intermediate cell and the microfilaments and microtubules in the light cell processes were observed to be well differentiated.

• Journal of Apiculture
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• v.34 no.3
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• pp.245-254
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• 2019

We investigated the anti-tumor effects and molecular mechanism of Brazil, China and Korean regional propolis on HeLa ovarian cancer cell line. Each propolis extracts was prepared by ethanol extraction method. Cytotoxicity of propolis extracts was determinated by EZ-cytox cell viability assay. To necessity of anti-tumor effect and molecular mechanism of propolis, we must be adjusting propolis concentration. Due to 100 ㎍/mL of propolis extract were reduced cell viability to less than 50%, we adjusted all of propolis concentration to 100 ㎍/mL. By Western blotting analysis, we confirmed that anti-tumor mechanism of Brazil, China and Korea regional propolis has significantly difference. All of propolis was activated apoptosis related molecules such as PARP, caspase-3. However, cell proliferation signaling molecules including Akt1, ERK and Bcl-2 were reduced the protein expression level. Especially, the expression of tumor suppressor protein p53 was significantly increased in propolis-treated group such as Gyeonggi, Chungbuk, Chungnam, Jeonbuk, Gyeongnam and China. The phosphorylation of Bax which as apoptosis indicator was appeared in propolis-treated group such as Gyeonggi, Gangwon, Chungnam, Gyeongbuk, China. In this results showed that the regional propolis has completely different mechanism in anti-tumor. Thus, propolis extracts may be useful source of functional materials on anti-cancer and it will be able to choose the suitable propolis for cancer therapy by analyzing individual characteristics.

• Journal of the Korea Convergence Society
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• v.10 no.4
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• pp.81-89
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• 2019

This study was analyzed about the relationship between culture microenvironment and cell differentiation of HPV 16 E6/E7-transfected immortalized oral keratinocyte(IHOK). By the alteration of culture environment, IHOK-EF and IHOK-EFKGM were obtained, and the modulation of cell properties was observed by cell proliferation assay, immunofluorescence, microarray, and quantitative real-time PCR analysis. IHOK-EF losed the properties of epithelial cells and obtained the properties of mesenchymal cells, and in the result of microarray analysis, genes related to the inhibition of differentiation such as IL6, TWIST1, and ID2 were highly expressed in IHOK-EF. When the culture environment was recovered to initial environment, these changes were recovered partially, presenting the return of genes involved in the inhibition of differentiation such as IL6, and ID2, particularly. This study will contribute to understand adjustment aspect for cell surviving according to the change of culture microenvironment in the study for determining the cell characteristic, and facilitate therapeutic approach for human disease by applying surviving study according to the change of cancer microenvironment.